In a study of 12 patients with recurrent glioblastoma multiforme the therapeutic potential of combining HSVtk and IL-2 using a bicistronic Moloney vector, was reported using retroviral VPC (25,31).

Rather than using a bicistronic vector carrying both HSVtk and IL-2 genes, we have delivered HSVtk via retroviral VPC and IL-2 via direct intramuscular plasmid injection so that the IL-2 is in circulation longer (>10 days as reported earlier) (9).

As shown in Figure 2B, mice that are homozygous for the targeted insertion of the conditional rtTA-IRES-EGFP bicistronic transgene at ROSA26 locus can be bred with double transgenic mice that carry a tissue-specific Cre transgene and a doxycycline-inducible rtTA-dependent tet-O-GENE.

As E6 and E7 are expressed together from a bicistronic mRNA, such approaches generally lead to the reactivation of both pRb and p53, and to the attainment of a growth-arrested state that resembles the replicative senescence that is achieved by primary cells after they have reached their normal life span in culture.

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